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Normal Phase Chromatography

Normal Phase Hydrophilic Interaction Chromatography

While reversed phase is the most widely used mode of retention in HPLC, very polar compounds are often not sufficiently retained in low percent organic, or even in 100% aqueous mobile phase. The order of elution in normal phase is opposite that found in RPC for the same mixture of compounds. Although non-polar organic mobile phases and a silica stationary phase were used traditionally in normal phase LC, today most normal phase separations are performed with aqueous-organic mobile phases and a more polar-bonded stationary phase. This mode of HPLC is now commonly referred to as HILIC, hydrophilic interaction chromatography.

By using an amide or amino bonded phase column, polar compounds can be retained by a normal phase or hydrophilic interaction chromatography (HILIC; see references) retention mechanism using a mobile phase mixture of acetonitrile and ammonium acetate buffer. In contrast to the retention behavior in reversed phase, in HILIC, solutes will be retained longer when increasing the percent acetonitrile.


  TSKgel Amide-80                    TSKgel NH2-100
Bonded Phase                                                                                                                                - Amide-80 bonded phase does
not react with reducing sugars.
- Anomer formation can be
prevented by raising mobile
phase temperature up to 80°C.                                                                                                                                                          
- NH2-100 bonded phase is more stable than conventional amino phases due to a special endcapping prior to introduction of aminoalkyl groups.
- Amino-bonded phase can react with a reducing sugar to form a Schiff base.                     
Detectors
Can be used with evaporative light scattering 
(ELS) and mass spec (MS) detectors
Mobile Phase Stable in 100% organic eluents
Solutes Retains very polar drug candidates and drug metabolites
Applications - saccharides
- polyols (polyalchols)
- drug metabolites
- peptides
- melanine & cyanuric acid
- oligonucleotides
- polar compounds in
- fermentation broth
cystosine 
- saccharides
- polyols (polyalchols)
- methotrexate polyglutamate derivatives
- pyridylaminated oligosaccharides
- water-soluble vitamins
- nucleic acid fragments

Literature:

 • Instruction Manual Annex for 2µm

 • Instruction Manual


Particle Size (mean):   2 µm
Pore Size (mean): 10 nm
Functional Group: carbamoyl
pH Range: 2.0 - 7.5
Max. temperature : 50 ºC
Surface Area (m2/g): 450

P/N
Description
Particle size
Housing material ID (mm) Length (cm)
0023454
TSKgel Amide-80 2µm
2 µm
Stainless Steel 2 5
0023455
TSKgel Amide-80 2µm
2 µm
Stainless Steel 2 10
0023456
TSKgel Amide-80 2µm
2 µm
Stainless Steel 2 15
0023457
TSKgel Amide-80 2µm
2 µm
Stainless Steel 3 5
0023458
TSKgel Amide-80 2µm
2 µm
Stainless Steel 3 10
0023459
TSKgel Amide-80 2µm
2 µm
Stainless Steel 3 15
0023460
TSKgel guardclumn Amide-80 2μm
2 µm 
Stainless Steel 2 1

Literature:

 • Catalog

 • HILIC Column Products Table

 • Instruction Manual


Particle Size (mean):   3 µm
Pore Size (mean): 10 nm
Functional Group: carbamoyl
pH Range: 2.0 - 7.5
Max. temperature : 50 ºC
Surface Area (m2/g): 450

P/N
Description
Particle size
Housing material ID (mm) Length (cm)
0021864
TSKgel Amide-80 3µm
3 µm
Stainless Steel 2 5
0021865
TSKgel Amide-80 3µm
3 µm
Stainless Steel 2 15
0022850
TSKgel Amide-80 3µm
3 µm
Stainless Steel 3 5
0022851
TSKgel Amide-80 3µm
3 µm
Stainless Steel 3 10
0022852
TSKgel Amide-80 3µm
3 µm
Stainless Steel 3 15
0021862
Guard Cartridge for 2 and 3mmID columns, 3pk
3 µm
Stainless Steel 2 1
0019308
Guard Cartridge Holder for P/N 0021862
 
Stainless Steel 2 1
0021866
TSKgel Amide-80 3µm
3 µm
Stainless Steel 4.6 5
0022849
TSKgel Amide-80 3µm
3 µm
Stainless Steel 4.6 10
0021867
TSKgel Amide-80 3µm
3 µm
Stainless Steel 4.6 15
0021863
Guard Cartridge for 4.6mmID columns, 3pk
3 µm
Stainless Steel 3.2 1.5
0019018
Guard Cartridge Holder for P/N 0021863
 
Stainless Steel 3.2 1.5

Literature:

 • Separation Report No. 55

 • Separation Report No. 79

 • Catalog

 • HILIC Column Products Table

 • Instruction Manual


Particle Size (mean):   5 and 10 µm
Pore Size (mean): 10 nm
Functional Group: carbamoyl
pH Range: 2.0 - 7.5
Max. temperature : 80 ºC
Surface Area (m2/g): 450

P/N
Description
Particle size
Housing material ID (mm) Length (cm)
0020009
TSKgel Amide-80
5 µm
Stainless Steel 1 5
0020010
TSKgel Amide-80
5 µm
Stainless Steel 1 10
0021486
TSKgel Amide-80
5 µm
Stainless Steel 1 15
0021487
TSKgel Amide-80
5 µm
Stainless Steel 1 25
0019694
TSKgel Amide-80
5 µm
Stainless Steel 2 5
0019695
TSKgel Amide-80
5 µm
Stainless Steel 2 10
0019696
TSKgel Amide-80
5 µm
Stainless Steel 2 15
0019697
TSKgel Amide-80                                   
5 µm
Stainless Steel 2 25
 0021941 Guard Cartridge for 2mm ID columns, 3 pk, Replaces P/N 0042140
5 µm
Stainless Steel 2 1
0019308
Guard Cartridge holder for P/N 0021941
 
Stainless Steel 2 1
0019532
TSKgel Amide-80
5 µm
Stainless Steel 4.6 5
0019533
TSKgel Amide-80
5 µm
Stainless Steel 4.6 10
0013071
TSKgel Amide-80
5 µm
Stainless Steel 4.6 25
0019021
Guard Column for 4.6mm ID columns
5 µm
Stainless Steel 4.6 1
0019010
Guard Column for 4.6mm ID columns, 3 pk 5 µm Stainless Steel 3.2 1.5
 0019018
Guard Cartridge holder for P/N 0019010
  Stainless Steel  3.2
1.5
0014459
TSKgel Amide-80
10 µm
Stainless Steel 7.8 30
0014460
TSKgel Amide-80
10 µm
Stainless Steel 21.5 30
0014461
Guard column for P/N 0014460
10 µm
Stainless Steel 21.5 7.5

Literature:

 • Catalog

 • HILIC Column Products Table

 • Instruction Manual

 • Instruction Manual for NH2-100 DC


Particle Size (mean):   3 µm
Pore Size (mean): 10 nm
Functional Group: alkylamine
pH Range: 2.0 - 7.5
Max. temperature: 50 ºC
Surface Area (m2/g): 450

P/N
Description
Particle size
Housing material ID (mm) Length (cm)
0021967
TSKgel NH2-100
3 µm
Stainless Steel 2 5
0021968
TSKgel NH2-100
3 µm
Stainless Steel 2 15
0021971
Guard Cartridge for 2mmID columns, 3pk
3 µm
Stainless Steel 2 1
0019308
Guard Cartridge Holder for P/N 21971
Stainless Steel 2 1
0021999
TSKgel NH2-100 DC
3 µm
Stainless Steel 4.6 5
0021969
TSKgel NH2-100
3 µm
Stainless Steel 4.6 5
0021970
TSKgel NH2-100
3 µm
Stainless Steel 4.6 15
0021972
Guard Cartridge for 4.6mmID columns, 3pk
3 µm
Stainless Steel 3.2 1.5
0019018
Guard Cartridge Holder for P/N 21972
Stainless Steel 3.2 1.5